Nitrous oxide (N 2 O) is one of the most important greenhouse gases and a major sink for stratospheric ozone. Estuaries are sites of intense biological production and N 2 O emissions. We aimed to identify hot spots of N 2 O production and potential pathways contributing to N 2 O concentrations in the surface water of the tidal Elbe estuary. During two research cruises in April and June 2015, surface water N 2 O concentrations were measured along the salinity gradient of the Elbe estuary by using a laser-based on-line analyzer coupled to an equilibrator. Based on these high-resolution N 2 O profiles, N 2 O saturations, and fluxes across the surface water/atmosphere interface were calculated. Additional measurements of DIN concentrations, oxygen concentration, and salinity were performed. Highest N 2 O concentrations were determined in the Hamburg port region reaching maximum values of 32.3 nM in April 2015 and 52.2 nM in June 2015. These results identify the Hamburg port region as a significant hot spot of N 2 O production, where linear correlations of AOU-N 2 O xs indicate nitrification as an important contributor to N 2 O production in the freshwater part. However, in the region with lowest oxygen saturation, sediment denitrification obviously affected water column N 2 O saturation. The average N 2 O saturation over the entire estuary was 201% (SD: ±94%), with an average estuarine N 2 O flux density of 48 µmol m −2 d −1 and an overall emission of 0.18 Gg N 2 O y −1 . In comparison to previous studies, our data indicate that N 2 O production pathways over the whole estuarine freshwater part have changed from predominant denitrification in the 1980s toward significant production from nitrification in the present estuary. Despite a significant reduction in N 2 O saturation compared to the 1980s, N 2 O concentrations nowadays remain on a high level, comparable to the mid-90s, although a steady decrease of DIN inputs occurred over the last decades. Hence, the Elbe estuary still remains an important source of N 2 O to the atmosphere.